Multimodal peptide ligand extracts parvovirus from interface in affinity aqueous two-phase system.

Area of Science:

• Biochemistry
• Biophysical Chemistry
• Separation Science

Background:

• Aqueous two-phase systems (ATPS) are utilized for bioseparation and microencapsulation, aiming to partition biomolecules into preferred phases.
• Understanding biomolecule behavior at the interface of these phases is crucial but often lacking.
• Tie-lines (TL) characterize ATPS, defining thermodynamic equilibrium and phase compositions (e.g., PEG-rich or droplet phases).

Purpose of the Study:

• To investigate porcine parvovirus (PPV) partitioning behavior within ATPS.
• To explore methods for improving PPV recovery by minimizing interfacial adsorption.
• To evaluate the impact of a PEG-peptide conjugate on PPV partitioning and interfacial behavior.

Main Methods:

• Utilized aqueous two-phase systems (ATPS) with varying tie-lines (TL) to study porcine parvovirus (PPV) partitioning.
• Formulated a PEG 10 kDa-peptide conjugate incorporating the WRW ligand.
• Analyzed PPV distribution between phases and at the interface under different system conditions (e.g., salt and PEG concentrations, TL).

Main Results:

• Higher PPV recovery was observed in PEG-rich bulk phases with citrate-rich droplets at high salt and PEG concentrations.
• The PEG-peptide conjugate significantly reduced PPV adsorption at the interface.
• The conjugate enhanced PPV recovery in lower TL systems (lower viscosity, reduced phase-forming component concentrations).

Conclusions:

• The PEG-peptide conjugate offers a strategy to improve virus recovery in ATPS, particularly in lower viscosity systems.
• This work provides insights into interfacial phenomena in ATPS and methods to direct biomolecule partitioning into phases rather than interfaces.
• The findings contribute to optimizing bioseparation techniques for viral recovery and purification.